Analyzing the role of heparan sulfate proteoglycans in axon guidance in vivo in zebrafish.
Analyzing the role of heparan sulfate proteoglycans in axon guidance in vivo in zebrafish.
复制标题
分析硫酸乙酰肝素蛋白聚糖在斑马鱼体内轴突引导中的作用。
DOI：
10.1007/978-1-4939-1714-3_36
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Poulain,FabienneE
中科院分区：
文献类型：
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作者：
Poulain,FabienneE
神经生物学领域最令人着迷的问题之一是了解神经元连接是如何正确形成的。在发育过程中，神经元延伸轴突，轴突由吸引和排斥的线索引导沿着确定的路径到达它们的大脑目标。这些指导因子大多受硫酸乙酰肝素蛋白多糖(HSPGs)的调节，HSPGs是一类细胞表面和细胞外核心蛋白，连接着硫酸乙酰肝素(HS)糖胺聚糖。HS糖链的独特多样性和结构复杂性以及核心蛋白质的多样性被认为是产生大脑连接所必需的复杂“糖码”。而HSPGs的功能已经得到了很好的描述。对于它们在脊椎动物神经系统发育中的作用，人们知之甚少。在本章中，我们描述了在斑马鱼体内直接研究HSPGs在轴突引导中的作用的优势和可用的不同方法。我们提供了在活体内显示轴突的方案，包括精确的染料标记和延时成像，以及干扰HS修饰酶和核心蛋白的功能，包括吗啡、DNA或RNA注射。
One of the most fascinating questions in the field of neurobiology is to understand how neuronal connections are properly formed. During development, neurons extend axons that are guided along defined paths by attractive and repulsive cues to reach their brain target. Most of these guidance factors are regulated by heparan sulfate proteoglycans (HSPGs), a family of cell-surface and extracellular core proteins with attached heparan sulfate (HS) glycosaminoglycans. The unique diversity and structural complexity of HS sugar chains, as well as the variety of core proteins, have been proposed to generate a complex “sugar code” essential for brain wiring. While the functions of HSPGs have been well characterized inC. elegansorDrosophila, relatively little is known about their roles in nervous system development in vertebrates. In this chapter, we describe the advantages and the different methods available to study the roles of HSPGs in axon guidance directly in vivo in zebrafish. We provide protocols for visualizing axons in vivo, including precise dye labeling and time-lapse imaging, and for disturbing the functions of HS-modifying enzymes and core proteins, including morpholino, DNA, or RNA injections.